AUTHOR=Munawaroh Heli Siti Halimatul , Apdila Egi Tritya , Awai Koichiro 

TITLE=hetN and patS Mutations Enhance Accumulation of Fatty Alcohols in the hglT Mutants of Anabaena sp. PCC 7120

JOURNAL=Frontiers in Plant Science

VOLUME=11

YEAR=2020

URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2020.00804

DOI=10.3389/fpls.2020.00804

ISSN=1664-462X

ABSTRACT=<p>The heterocysts present in filamentous cyanobacteria such as <italic>Anabaena</italic> sp. PCC 7120 are known to be regulated by HetN and PatS, the repressors of heterocyst differentiation; therefore, the inactivation of these proteins will result in the formation of multiple heterocysts. To enhance the accumulation of fatty alcohols synthesized in the heterocyst, we introduced mutations of these repressors to increase heterocyst frequency. First, we isolated double mutants of <italic>hetN</italic> and <italic>patS</italic> and confirmed that the null mutation of these genes promoted higher frequencies of heterocyst formation and higher accumulation of heterocyst-specific glycolipids (Hgls) compared with its wild type. Next, we combined <italic>hetN</italic> and <italic>patS</italic> mutations with an <italic>hglT</italic> (encoding glycosyltransferase, an enzyme involved in Hgl synthesis) mutation to increase the accumulation of fatty alcohols since knockout mutation of <italic>hglT</italic> results in accumulation of very long chain fatty alcohol, the precursor of Hgl. We also observed retarded growth, lower chlorophyll content and up to a five-fold decrease in photosynthetic activity of the <italic>hetN</italic>/<italic>patS</italic>/<italic>hglT</italic> triple mutants. In contrast, the triple mutants showed three times higher heterocyst formation frequencies than the <italic>hglT</italic> single mutant and wild type. The production rate of fatty alcohol in the triple mutants attained a value 1.41 nmol/mL OD<sub>730</sub>, whereas accumulation of Hgls in the wild type was 0.90 nmol/mL OD<sub>730</sub>. Aeration of culture improved the accumulation of fatty alcohols in <italic>hetN</italic>/<italic>patS</italic>/<italic>hglT</italic> mutant cells up to 2.97 nmol/mL OD<sub>730</sub> compared with cells cultured by rotation. Our study outlines an alternative strategy for fatty alcohol production supported by photosynthesis and nitrogen fixation.</p>